The present invention relates to the fields of molecular biology and nucleic acid analysis. More specifically, the present invention relates to an improved method for attaching a desired compound, preferably a nucleic acid, to a silaceous or silane-containing substrate, such as a glass, e.g., channel glass, glass slide, quartz, SiO2, porous silica, micromachined silicon, oxidized silicon, or other silanized material, or a solid material coated with any of the above. The invention provides an efficient method for achieving the stable attachment of a desired compound to a silane-containing substrate that obviates the need to derivatize or activate such silane-containing substrate to facilitate attachment.
More particularly, the present invention relates to a method for stably attaching a compound having at least one amine and at least one hydroxyl functional group, e.g., an aminopropanol containing compound, to a silaceous or silane-containing substrate, e.g., plain (underivatized) glass, porous silica, or oxidized silicon. The resultant silane-containing substrate, to which is stably attached said functional group-containing compound, can be used in any application wherein compounds immobilized to silane-containing substrates, e.g., glass (e.g., channel glass, glass slide), quartz, SiO2 porous silica, micromachined silicon, or oxidized silicon materials, or materials coated with the foregoing, are useful. Such methods include, in particular, hybridization, nucleic acid isolation, nucleic acid purification, immunoassay, and immunopurification methods.
The present invention provides a method for attaching a compound having at least one amine group and at least one hydroxyl group, to a silane-containing substrate, e.g., a glass, porous silica, oxidized silicon, or other silaceous or silane-containing material. More specifically, the present invention provides a method for attaching an aminopropanol-containing compound to a silane-containing substrate, e.g., a glass, porous silica, oxidized silicon, or a silanized material.
Unlike conventional methods wherein a desired compound is attached to a silaceous or silane-containing substrate, such as glass or porous silica, there is no need in the present invention to derivatize or activate the substrate, e.g., glass (such as previous xe2x80x9cepoxysilane activation methodxe2x80x9d) to facilitate attachment. Rather, as discussed in detail infra, the subject invention instead provides a method for the stable attachment of a compound containing at least one amino and hydroxyl functional group to plain, i.e., underivatized, or non-activated glass, porous silica, or oxidized silicon. As discussed in detail infra, both of such groups are involved in the attachment, and are preferably proximate to one another, e.g., within 1 to 6 atoms of one another, and more preferably are comprised on adjacent carbon atoms. This method is particularly useful for attaching derivatized oligonucleotides, in particular 3xe2x80x2 or 5xe2x80x2 aminopropanol-derivatized oligonucleotides, to silaceous or silane-containing substrates such as glasses, porous silicas, and oxidized silicon.
The invention provides improved methods for stably attaching desired compounds to a silaceous or silane-containing substrate, e.g., glass (e.g., channel glass, glass slide), porous silica, micromachined silicon, or oxidized silicon material, or a material coated with any of the foregoing.
The invention more specifically provides an efficient method for stably attaching a compound having at least one amine and at least one hydroxyl group, which functional groups provide for the stable attachment of such compound to a silaceous or silane-containing substrate, e.g., a glass, such as channel glass, glass slide, quartz, SiO2, micromachined silicon, porous silica, or oxidized silicon material.
The invention specifically provides an efficient method for stably attaching an aminopropanol-containing compound to a silaceous o r silane-containing substrate, e.g., a glass, porous silica, micromachined silicon, quartz, SiO2, silanized material, or oxidized silicon material.
The invention even more specifically provides an efficient method for stably attaching a desired nucleic acid, e.g., an oligonucleotide, to a silane-containing substrate, e.g., a glass, porous silica, or oxidized silicon, by the following steps:
(i) obtaining a derivatized nucleic acid, e.g., an oligonucleotide, that comprises at least one amine and at least one hydroxyl group (wherein said functional groups are distinguished from the NH2xe2x80x94CO bonds that provide NH2 functional moieties in base-paired DNA""s) which functional groups facilitate the attachment of said nucleic acid to a silane-containing substrate; and
(ii) contacting said derivatized nucleic acid containing said at least one amine and hydroxyl group with a silaceous or silane-containing substrate, e.g., plain glass, porous silica, or oxidized silicon material under conditions that provide for the stable attachment of such compound to said silane-containing substrate, e.g., plain (underivatized) glass, channel glass, glass slide, porous silica, micromachined silicon, quartz, SiO2, silanized material, oxidized silicon, or a material coated with any of the foregoing.
The invention also specifically provides a method for stably attaching an aminopropanol derivatized nucleic acid, preferably a 3xe2x80x2- or 5xe2x80x2-aminopropanol derivatized oligonucleotide to a silaceous or silane-containing substrate by the following steps:
(i) obtaining an aminopropanol-derivatized nucleic acid, e.g., a 3xe2x80x2- or 5xe2x80x2-aminopropanol derivatized oligonucleotide; and
(ii) contacting said aminopropanol-derivatized nucleic acid with a silaceous or silane-containing substrate, e.g., plain glass, porous silica, or oxidized silicon, material, under conditions that provide for the stable attachment of such derivatized nucleic acid to said silane-containing substrate, e.g., plain glass or oxidized silicon.
The invention also provides oligonucleotide arrays that are stably attached to silaceous or silane-containing substrates, e.g., plain (underivatized) glass, or oxidized silicon.
The invention also provides uses of oligonucleotide that are stably attached in arrays to silane-containing substrates, e.g., plain (underivatized) glass, or oxidized silicon, for detection and/or quantification of target nucleic acid sequences.
The invention also specifically provides stable oligonucleotide which comprise 3xe2x80x2-aminopropanol-derivatized oligonucleotides that are stably attached to plain (underivatized) glasses, such as microchannel glass, porous silica, or oxidized silicon materials, and method to use these stably oligonucleotide arrays for detecting target nucleic acids having a complementary sequence.